Joshi, P and Mondal, PP (2022) Single-molecule clustering for super-resolution optical fluorescence microscopy. In: Photonics, 9 (1).
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Abstract
Molecular assembly in a complex cellular environment is vital for understanding underlying biological mechanisms. Biophysical parameters (such as single-molecule cluster density, cluster-area, pairwise distance, and number of molecules per cluster) related to molecular clusters directly associate with the physiological state (healthy/diseased) of a cell. Using super-resolution imaging along with powerful clustering methods (K-means, Gaussian mixture, and point clustering), we estimated these critical biophysical parameters associated with dense and sparse molecular clusters. We investigated Hemaglutinin (HA) molecules in an Influenza type A disease model. Sub-sequently, clustering parameters were estimated for transfected NIH3T3 cells. Investigations on test sample (randomly generated clusters) and NIH3T3 cells (expressing Dendra2-Hemaglutinin (Dendra2-HA) photoactivable molecules) show a significant disparity among the existing clustering techniques. It is observed that a single method is inadequate for estimating all relevant biophysical parameters accurately. Thus, a multimodel approach is necessary in order to characterize molecular clusters and determine critical parameters. The proposed study involving optical system development, photoactivable sample synthesis, and advanced clustering methods may facilitate a better understanding of single molecular clusters. Potential applications are in the emerging field of cell biology, biophysics, and fluorescence imaging. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
| Item Type: | Journal Article |
|---|---|
| Publication: | Photonics |
| Publisher: | MDPI |
| Additional Information: | The copyright for this article belongs to the Author. |
| Department/Centre: | Division of Physical & Mathematical Sciences > Instrumentation Appiled Physics |
| Date Deposited: | 12 Jan 2022 05:45 |
| Last Modified: | 12 Jan 2022 05:45 |
| URI: | http://eprints.iisc.ac.in/id/eprint/70923 |
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